The Rapid Single Spore Enumeration Assay (RapidSSEA) is being developed for the Mars Technology Program to aid planetary protection personnel in their efforts to validate bioburden reduction on spacecraft surfaces prior to launch. RapidSSEA is based on imaging and counting individual bacterial spores in a microscope field-of-view, where the contrast is generated by a highly luminescent complex that forms when dipicolinic acid (DPA) is released from spores during germination and binds to terbium (Tb/sup 3+/) ions in the surrounding medium. Under pulsed UV excitation, the long-lived (/spl tau/ /spl sim/ 1 ms) DPA-triggered Tb luminescence is imaged by a lifetime-gated camera, essentially eliminating fluorescent interferents from the image, which enables counting of bacterial spores swabbed from environmental samples. Here we report results from the first side-by-side comparison of RapidSSEA and the NASA standard assay. Both assays were used to assess the spore population on stainless steel coupons inoculated with Bacillus atrophaeus spores. Using RapidSSEA, 38.0% /spl plusmn/ 5.8% of the total inoculated spore population was recovered and counted within 30 minutes.
A Rapid Single Spore Enumeration Assay
2006 IEEE Aerospace Conference ; 13 pp.
2006-01-01
1063873 byte
Conference paper
Electronic Resource
English
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